Nitrogenase from the photosynthetic bacterium Rhodopseudomonas capsulata: purification and molecular properties.
Nitrogenase proteins were isolated from cultures of the photosynthetic bacterium Rhodopseudomonas capsulata grown on a limiting amount of ammonia. Under these conditions, the nitrogenase N2ase A was active in vivo, and nitrogenase activity in vitro was not dependent upon manganese and the activating factor. The nitrogenase proteins were also isolated from nitrogen-limited cultures in which the in vivo nitrogenase activity had been stopped by an ammonia shock. This nitrogenase activity, N2ase R, showed an in vitro requirement for manganese and the activating factor for maximal activity. The Mo-Fe protein (dinitrogenase) was composed of two dissimilar subunits with molecular weights of 55,000 and 59,500; the Fe protein (dinitrogenase reductase), from either type of culture, was composed of a single subunit (molecular weight), 33,500). The metal and acid labile sulfur contents of both nitrogenase proteins were similar to those found for previously isolated nitrogenases. The Fe proteins from both N2ase A and N2ase R contained phosphate and ribose, 2 mol of each per mol of N2ase R Fe protein and about 1 mol of each per mol of N2ase A Fe protein. The greatest difference between the two types of Fe protein was that the N2ase R Fe protein contained about 1 mol per mol of an adenine-like molecule, whereas the N2ase A Fe protein content of this compound was insignificant. These results are compared with various models previously presented for the short-term regulation of nitrogenase activity in the photosynthetic bacteria.
D000225:Adenine / Q000032:analysis; D000596:Amino Acids / Q000032:analysis; D000645:Ammonium Sulfate / Q000494:pharmacology; D007526:Isoelectric Point; D008345:Manganese / Q000494:pharmacology; D008970:Molecular Weight; D009591:Nitrogenase / Q000032:analysis / Q000302:isolation & purification* / Q000378:metabolism; D010710:Phosphates / Q000032:analysis; D012241:Rhodopseudomonas / Q000201:enzymology*; D012266:Ribose / Q000032:analysis
D000596:Amino Acids; D010710:Phosphates; D008345:Manganese; D012266:Ribose; D009591:Nitrogenase; D000225:Adenine; D000645:Ammonium Sulfate